Establishing a physiological environment for visualized in vitro brain slice recordings by increasing oxygen supply and modifying aCSF content

Our insights into the basic characteristics of neuronal function were significantly advanced by combining the in vitro slice technique with the visualization of neurons and their processes. The visualization through water immersion objectives requires keeping slices submerged in recording chambers where delivering artificial cerebro-spinal fluid (aCSF) at flow rates of 2–3 ml/min results in a limited oxygen supply [Hájos N, Ellender TJ, Zemankovics R, Mann EO, Exley R, Cragg SJ, et al. Maintaining network activity in submerged hippocampal slices: importance of oxygen supply. Eur J Neurosci 2009;29:319–27]. Here we review two methods aimed at providing sufficient oxygen levels to neurons in submerged slices to enable high energy consuming processes such as elevated firing rates or network oscillations. The use of these methods may also influence the outcome of other electrophysiological experiments in submerged slices including the study of intercellular signaling pathways. In addition, we also emphasize the importance of various aCSF constituents used in in vitro experiments.